Polymorphisms of HM30181 mesylate

ABSTRACT

HM30181 (shown below) can be used to improve absorption of cancer chemotherapy drugs, such as paclitaxel. Herein are described various polymorphisms of HM30181, in particular polymorphism C, as well as their physical properties and methods for their preparation and characterization.

This application claims the benefit of U.S. Provisional PatentApplication No. 63/107,720 filed on Oct. 30, 2020, and ProvisionalPatent Application No. 63/107,792 filed on Oct. 30, 2020. These and allother referenced extrinsic materials are incorporated herein byreference in their entirety. Where a definition or use of a term in areference that is incorporated by reference is inconsistent or contraryto the definition of that term provided herein, the definition of thatterm provided herein is deemed to be controlling.

FIELD OF THE INVENTION

The field of the invention is P-glycoprotein inhibitors, in particularHM30181 mesylate.

BACKGROUND

The background description includes information that may be useful inunderstanding the present invention. It is not an admission that any ofthe information provided herein is prior art or relevant to thepresently claimed invention, or that any publication specifically orimplicitly referenced is prior art.

P-glycoprotein (P-gp) is an ATP dependent efflux pump protein with awide range of substrate specificity that is found throughout the majorgenera. Due to its broad distribution and its function it is thought tobe a defense mechanism that actively transport toxins out of cells. Inhumans P-gp can transport substrate compounds from intestinal epithelialcells back into the intestinal lumen, from the blood brain barrier backinto adjacent capillaries, from the proximal tubule of the kidney intothe urinary filtrate, and from liver cells into the bile ducts.

Unfortunately, a number of drugs utilized in chemotherapy are substratesfor P-gp. P-gp activity, therefore, can reduce bioavailability andeffectiveness of chemotherapeutic drugs. In such instancesadministration of P-gp inhibitors can be useful in improving theresponse to chemotherapy. Accordingly, over the last 30 years a numberof pharmaceutically useful P-gp inhibitors (such as amiodarone,clarithromycin, cyclosporin, colchicine, diltiazem, erythromycin,felodipine, ketoconazole, lansoprazole, omeprazole, nifedipine,paroxetine, reserpine, saquinavir, sertraline, quinidine, tamoxifen,verapamil, and duloxetine) have been developed.

HM30181 mesylate is a third generation P-gp inhibitor that has beenstudied for use with paclitaxel. HM30181 mesylate selectively inhibitsP-gp in the intestinal epithelium, improving absorption of orallyadministered chemotherapeutic drugs without increasing potentiallydetrimental transport across the blood-brain barrier. The structure ofHM30181 mesylate is shown below.

The pharmacokinetics, bioavailability, and incidence of side effects oforally administered HM30181 mesylate are less than optimal, however.

Thus, there is still a need for polymorphisms of HM30181 mesylate thatcan provide improved absorption, improved pharmacokinetics, and/orreduced side effects upon administration.

SUMMARY OF THE INVENTION

The inventive subject matter provides polymorphisms of HM31081 mesylate,methods for their preparation and characterization, and methods fortheir use.

One embodiment of the inventive concept is a composition that includes acrystalline or partially crystalline form of HM30181 mesylate, where thecrystalline or partially crystalline form includes polymorph B,polymorph C, polymorph D, polymorph E, polymorph F, polymorph G,polymorph H, polymorph I, polymorph J, polymorph K, polymorph L,polymorph M, and/or polymorph N. In some of such embodiments thecrystalline or partially crystalline form is polymorph B, and has anX-ray diffraction pattern corresponding to FIG. 40 and an endotherm atabout 159.92° C. In some of such embodiments the crystalline orpartially crystalline form is polymorph Type C, and has an X-raydiffraction pattern corresponding to FIG. 42 and an endotherm at about159.6° C. In some of such embodiments the crystalline or partiallycrystalline form is polymorph Type C, and the crystalline form has anX-ray diffraction pattern with 2Theta maxima at about 6.4° and about8.0°. In some of such embodiments the crystalline or partiallycrystalline form is polymorph Type C, and has a unit cell with a volumeof about 1180 Å³, where a is about 7 Å, b is about 15 Å, c is about 18Å, alpha is about 52°, beta is about 62°, and gamma is about 90°. Insome of such embodiments the crystalline or partially crystalline formis polymorph C, and has an X-ray diffraction pattern corresponding toFIG. 42 and an endotherm at about 159.60° C., and can be a monohydrate.In some of such embodiments the crystalline or partially crystallineform is polymorph D, and has an X-ray diffraction pattern correspondingto FIG. 45 and an endotherm at about 66.97° C. In some of suchembodiments the crystalline or partially crystalline form is polymorphE, and has an X-ray diffraction pattern corresponding to FIG. 47 and anendotherm at about 154.42° C., and can include DMA. In some of suchembodiments the crystalline or partially crystalline form is polymorphType E, and has an X-ray diffraction pattern corresponding to FIG. 47and an endotherm at about 154.4° C. In some of such embodiments thecrystalline or partially crystalline form is polymorph Type E, and hasan X-ray diffraction pattern comprising 2Theta maxima at about 4.2°,about 10.4°, about 10.7°, about 14.7°, about 16.8°, about 21°, about23.8°, about 26.6°, and about 27.7°. In some of such embodiments thecrystalline or partially crystalline form is polymorph Type E, and has aunit cell with a volume of about 1620 Å³, where a is about 8 Å, b isabout 10 Å, c is about 24 Å, alpha is about 75°, beta is about 80°, andgamma is about 110°. In some of such embodiments the crystalline orpartially crystalline form is polymorph F, and has an X-ray diffractionpattern corresponding to FIG. 50 has an endotherm at about 148.41° C.,and can include DMF. In some of such embodiments the crystalline orpartially crystalline form is polymorph G, and has an X-ray diffractionpattern corresponding to FIG. 53 and an endotherm at about 69.02° C. Insome of such embodiments the crystalline or partially crystalline formis polymorph H, and has an X-ray diffraction pattern corresponding toFIG. 55 and an endotherm at about 126.52° C. In some of such embodimentsthe crystalline or partially crystalline form is polymorph I, and has anX-ray diffraction pattern corresponding to FIG. 57 . In some of suchembodiments the crystalline or partially crystalline form is polymorphJ, and has an X-ray diffraction pattern corresponding to FIG. 58 . Insome of such embodiments the crystalline or partially crystalline formis polymorph K, and has an X-ray diffraction pattern corresponding toFIG. 60 . In some of such embodiments the crystalline or partiallycrystalline form is polymorph L, and has an X-ray diffraction patterncorresponding to FIG. 61 . In some of such embodiments the crystallineor partially crystalline form is polymorph M, and has an X-raydiffraction pattern corresponding to FIG. 62 . In some of suchembodiments the crystalline or partially crystalline form is polymorphN, has an X-ray diffraction pattern corresponding to FIG. 63 and anendotherms at about 159° C. and about 188° C., and can include methanol.

Another embodiment of the inventive concept is a method of inhibitingP-glycoprotein activity, by contacting P-glycoprotein with at least onecrystalline or partially crystalline form of HM30181 mesylate asdescribed above in an amount that is effective in inhibiting an activityof P-glycoprotein.

Another embodiment of the inventive concept is a method of treatingcancer by administering a chemotherapeutic drug that is a P-glycoproteinsubstrate to an individual that in need of treatment for cancer and alsoadministering a polymorph of HM30181 mesylate as described above in anamount that is effective to inhibit P-glycoprotein activity in theindividual.

Another embodiment of the inventive concept is the use of a polymorph ofHM30181 mesylate as described above in preparing a medicament fortreating cancer. Such a medicament can also include a chemotherapeuticdrug that is a P-glycoprotein substrate.

Another embodiment of the inventive concept is a formulation thatincludes a polymorph of HM30181 mesylate as described above and atherapeutic drug, where the therapeutic drug is a P-glycoproteinsubstrate. Such a therapeutic drug can be a chemotherapeutic drug thatis used in the treatment of cancer.

Various objects, features, aspects and advantages of the inventivesubject matter will become more apparent from the following detaileddescription of preferred embodiments, along with the accompanyingdrawing figures in which like numerals represent like components.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 : FIG. 1 shows an overlay of X-Ray Powder Diffraction (XRPD)results from HM30181 mesylate polymorphisms types A to N.

FIG. 2 : FIG. 2 shows results from XRPD studies of HM30181 mesylate TypeA starting material

FIG. 3 : FIG. 3 shows an overlay of results of DSC/TGA studies ofHM30181 mesylate Type A.

FIG. 4 : FIG. 4 shows results of ¹H-NMR studies of HM30181 mesylate TypeA.

FIG. 5 : FIG. 5 shows results of DVS of HM30181 mesylate Type A startingmaterial.

FIG. 6 : FIG. 6 shows an overlay of overlay of XRPD studies of Type Astarting material post-DVS.

FIG. 7 : FIG. 7 shows an overlay of results from Cyclic DSC studies ofHM30181 mesylate Type A and HM30181 mesylate Type A heated to 110° C.

FIG. 8 : FIG. 8 shows an overlay of results of XRPD studies of HM30181mesylate Type A and HM30181 mesylate Type A heated to 110° C.

FIG. 9 : FIG. 9 shows an overlay of cyclic DSC studies of HM30181mesylate Type A starting material and HM30181 mesylate Type A heated to185° C.

FIG. 10 : FIG. 10 shows an overlay of XRPD studies of HM30181 mesylateType A starting material and HM30181 mesylate Type A heated to 185° C.

FIG. 11 : FIG. 11 shows an overlay of results of cyclic DSC studies ofHM30181 mesylate Type A starting material and HM30181 mesylate Type Aheated to 200° C.

FIG. 12 : FIG. 12 shows an overlay of results of XRPD studies of HM30181mesylate Type A starting material and HM30181 mesylate Type A heated to200° C.

FIG. 13 : FIG. 13 shows results of XRPD studies of freebase HM3018-A

FIG. 14 : FIG. 14 shows an overlay of results of ¹H-NMR studies ofHM30181 mesylate Type A heated to 200° C., freebase HM3018-A, andHM30181 mesylate Type A starting material.

FIG. 15 : FIG. 15 shows an overlay of results of XRPD studies of HM30181mesylate Type B from slurry experiments.

FIG. 16 : FIG. 16 shows an overlay of results of XRPD studies of HM30181mesylate Type C from slurry experiments.

FIG. 17 : FIG. 17 shows an overlay of results of XRPD studies of HM30181mesylate Type D from slurry experiments.

FIG. 18 : FIG. 18 shows an overlay of results of XRPD studies of HM30181mesylate Type E from slurry experiments.

FIG. 19 : FIG. 19 shows an overlay of results of XRPD of HM30181mesylate Type F from slurry experiments.

FIG. 20 : FIG. 20 shows an overlay of results of XRPD studies of HM30181mesylate Type G from slurry experiments.

FIG. 21 : FIG. 21 shows an overlay of results of XRPD studies of HM30181mesylate Type D from liquid vapor diffusion experiments.

FIG. 22 : FIG. 22 shows an overlay of results of XRPD studies of HM30181mesylate Type E from liquid vapor diffusion experiments.

FIG. 23 : FIG. 23 shows an overlay of results of XRPD studies of HM30181mesylate Type H from liquid vapor diffusion experiments.

FIG. 24 : FIG. 24 shows an overlay of results of XRPD studies of HM30181mesylate Type I and HM30181 mesylate Type J from liquid vapor diffusionexperiments.

FIG. 25 : FIG. 25 shows an overlay of results of XRPD studies of HM30181mesylate Type C polymorph generated by cooling.

FIG. 26 : FIG. 26 shows an overlay of results of XRPD studies of HM30181mesylate Type C with samples from anti-solvent experiments.

FIG. 27 : FIG. 27 shows an overlay of results of XRPD studies of HM30181mesylate Type F with sample from anti-solvent experiments.

FIG. 28 : FIG. 28 shows an overlay of results of XRPD studies of HM30181mesylate Type J with samples from anti-solvent experiments.

FIG. 29 : FIG. 29 shows an overlay of results of XRPD studies of HM30181mesylate Type K from anti-solvent experiments.

FIG. 30 : FIG. 30 shows an overlay of results of XRPD studies of HM30181mesylate Type L from anti-solvent experiments.

FIG. 31 : FIG. 31 shows an overlay of results of XRPD studies of HM30181mesylate Type M from anti-solvent experiments.

FIG. 32 : FIG. 32 shows an overlay of results of XRPD studies of HM30181mesylate Type B large scale studies.

FIG. 33 : FIG. 33 shows an overlay of results of XRPD studies of HM30181mesylate Type C large scale studies.

FIG. 34 : FIG. 34 shows an overlay of XRPD studies of HM30181 mesylateType D large scale studies.

FIG. 35 : FIG. 35 shows an overlay of results of XRPD studies of HM30181mesylate Type E large scale studies.

FIG. 36 : FIG. 36 shows an overlay of results of XRPD studies of HM30181mesylate Type G large scale studies.

FIG. 37 : FIG. 37 shows an overlay of results of XRPD studies of HM30181mesylate Type F and HM30181 mesylate Type G large scale studies.

FIG. 38 : FIG. 38 shows an overlay of results of ¹H-NMR studies ofHM30181 mesylate Type G large scale studies.

FIG. 39 : FIG. 39 shows an overlay of results of ¹H-NMR studies HM30181mesylate polymorphisms large scale studies.

FIG. 40 : FIG. 40 shows results of XRPD studies of HM30181 mesylate TypeB.

FIG. 41 : FIG. 41 shows an overlay of DSC and TGA results from HM30181mesylate Type B.

FIG. 42 : FIG. 42 shows results of XRPD of HM30181 mesylate Type C.

FIG. 43 : FIG. 43 shows an overlay of DSC and TGA results of HM30181mesylate Type C.

FIG. 44 : FIG. 44 shows results of ¹H-NMR of HM30181 mesylate Type C.

FIG. 45 : FIG. 45 shows results of XRPD of HM30181 mesylate Type D.

FIG. 46 : FIG. 46 shows an overlay of DSC and TGA results from HM30181mesylate Type D.

FIG. 47 : FIG. 47 shows results of XRPD of HM30181 mesylate Type E.

FIG. 48 : FIG. 48 shows an overlay of results from DSC and TGA ofHM30181 mesylate Type E.

FIG. 49 : FIG. 49 shows results of ¹H-NMR of HM30181 mesylate Type E.

FIG. 50 : FIG. 50 shows results from XRPD of HM30181 mesylate Type F.

FIG. 51 : FIG. 51 shows an overlay of results from DSC and TGA ofHM30181 mesylate Type F.

FIG. 52 : FIG. 52 shows results from ¹H-NMR of HM30181 mesylate Type F

FIG. 53 : FIG. 53 shows results from XRPD of HM30181 mesylate Type G.

FIG. 54 : FIG. 54 shows an overlay of results from DSC and TGA ofHM30181 mesylate Type G.

FIG. 55 : FIG. 55 shows results from XRPD of HM30181 mesylate Type H.

FIG. 56 : FIG. 56 shows an overlay of results from DSC and TGA ofHM30181 mesylate Type H.

FIG. 57 : FIG. 57 shows results from XRPD of HM30181 mesylate Type I.

FIG. 58 : FIG. 58 shows results from XRPD of HM30181 mesylate Type J.

FIG. 59 : FIG. 59 shows results from TGA of HM30181 mesylate Type J.

FIG. 60 : FIG. 60 shows results from XRPD of HM30181 mesylate Type K.

FIG. 61 : FIG. 61 shows results from XRPD of HM30181 mesylate Type L.

FIG. 62 : FIG. 62 shows results from XRPD of HM30181 mesylate Type M.

FIG. 63 : FIG. 63 shows results from XRPD of HM30181 mesylate Type N.

FIG. 64 : FIG. 64 shows an overlay of DSC and TGA results from HM30181mesylate Type N.

FIG. 65 : FIG. 65 shows results of ¹H-NMR of HM30181 mesylate Type N.

DETAILED DESCRIPTION

The following description includes information that may be useful inunderstanding the present invention. It is not an admission that any ofthe information provided herein is prior art or relevant to thepresently claimed invention, or that any publication specifically orimplicitly referenced is prior art.

The inventive subject matter provides a wide range of polymorphisms ofHM30181 mesylate and methods for their preparation. The variouspolymorphisms are shown to be structurally distinct by X-ray diffractionand various physical properties. Polymorphs of HM30181 mesylate withimproved pharmacokinetics, reduced incidence of side effects, reduceddosing schedules, etc. can be identified among these by conventionalmethods (e.g., animal studies, clinical studies, etc.).

One should appreciate that the disclosed techniques provide manyadvantageous technical effects including improving absorption ofchemotherapeutic drugs while maintaining patency of the blood-brainbarrier and reducing the incidence of developing drug resistance duringcancer treatment.

The following discussion provides many example embodiments of theinventive subject matter. Although each embodiment represents a singlecombination of inventive elements, the inventive subject matter isconsidered to include all possible combinations of the disclosedelements. Thus if one embodiment comprises elements A, B, and C, and asecond embodiment comprises elements B and D, then the inventive subjectmatter is also considered to include other remaining combinations of A,B, C, or D, even if not explicitly disclosed.

In some embodiments, the numbers expressing quantities of ingredients,properties such as concentration, reaction conditions, and so forth,used to describe and claim certain embodiments of the invention are tobe understood as being modified in some instances by the term “about.”Accordingly, in some embodiments, the numerical parameters set forth inthe written description and attached claims are approximations that canvary depending upon the desired properties sought to be obtained by aparticular embodiment. In some embodiments, the numerical parametersshould be construed in light of the number of reported significantdigits and by applying ordinary rounding techniques. Notwithstandingthat the numerical ranges and parameters setting forth the broad scopeof some embodiments of the invention are approximations, the numericalvalues set forth in the specific examples are reported as precisely aspracticable. The numerical values presented in some embodiments of theinvention may contain certain errors necessarily resulting from thestandard deviation found in their respective testing measurements.

As used in the description herein and throughout the claims that follow,the meaning of “a,” “an,” and “the” includes plural reference unless thecontext clearly dictates otherwise. Also, as used in the descriptionherein, the meaning of “in” includes “in” and “on” unless the contextclearly dictates otherwise.

The recitation of ranges of values herein is merely intended to serve asa shorthand method of referring individually to each separate valuefalling within the range. Unless otherwise indicated herein, eachindividual value is incorporated into the specification as if it wereindividually recited herein. All methods described herein can beperformed in any suitable order unless otherwise indicated herein orotherwise clearly contradicted by context. The use of any and allexamples, or exemplary language (e.g. “such as”) provided with respectto certain embodiments herein is intended merely to better illuminatethe invention and does not pose a limitation on the scope of theinvention otherwise claimed. No language in the specification should beconstrued as indicating any non-claimed element essential to thepractice of the invention.

Groupings of alternative elements or embodiments of the inventiondisclosed herein are not to be construed as limitations. Each groupmember can be referred to and claimed individually or in any combinationwith other members of the group or other elements found herein. One ormore members of a group can be included in, or deleted from, a group forreasons of convenience and/or patentability. When any such inclusion ordeletion occurs, the specification is herein deemed to contain the groupas modified thus fulfilling the written description of all Markushgroups used in the appended claims.

Inventors have identified 1a number of polymorphisms of HM30181mesylate, including Type A to Type N (see Table 1). Some of thepolymorphisms can include metastable solvates. The Inventors believethat one or more of these polymorphs can have improved pharmacokineticsand/or bioavailability relative to prior art formulations of HM30181mesylate. Inventors believe that such improvements can permit the use oflower doses that reduce or eliminate side effects associated withtreatment using prior art formulations of HM30181 mesylate.

TABLE 0 Crystal DSC form endotherm Batch (XRPD) (° C.) TGA weight lossIdentification B00505-05-C Type A 181.41 2.6% before 150° C. Monohydrate(hygroscopic (starting material) to 2.26% at 95% RH)6007235-16-A1-airdry Type B 159.92 1.9% before 170° C. Solvate/hydrate6004273-10-C-vd Type C 159.60 3.0% before 175° C. Monohydrate6007235-16-A21-airdry Type D 66.97 (peak), 14.71% before 150° C.Solvate/hydrate 114 (exotherm) 6004273-10-E-vd Type E 154.42 5.247%before 168° C. DMA solvate 6004273-10-G-vd Type F 148.41 5.123% weightloss DMF solvate before 180° C. 6007235-16-B26-airdry Type G  69.021.451% before 150° C. Solvate/hydrate 233.29 6007235-19-B10 Type H126.52 7.444% weight loss Solvate/hydrate before 200° C. 6007235-19-E3Type I NA NA Weak solvate- converts to Type J on air-drying6007235-19-E5 Type J NA 2.887% weight loss Solvate/hydrate before 150°C. 6004273-06-A4 Type K Insufficient Insufficient sample Possiblesolvate/hydrate sample 6004273-06-A60 Type L Insufficient Insufficientsample Possible solvate/hydrate sample 6004273-06-A56 Type MInsufficient Insufficient sample Possible solvate/hydrate sample6004273-10-B-vd Type N 159.28 2.128% weight loss Possible MeOH 188.47before 180° C. solvate/hydrateTable 1FIG. 1 provides a summary overlay of X-Ray Powder Diffraction (XRPD)results from HM30181 mesylate polymorphisms types A to N, as provided inTable 1.

A prior art HM30181 mesylate salt monohydrate material (i.e., a startingmaterial) can be synthesized as described in PCT application publicationnumber WO 2005/033097 or U.S. Pat. No. 9,283,218 which are incorporatedherein by reference. This starting material was characterized by XRPD,TGA, DSC, and DVS (see below), and was identified as crystalline Type Aby XRPD (see FIG. 2 ).

By DSC, HM30181 mesylate Type A displayed an endotherm at 181.41° C.(see FIG. 3 ). By TGA, HM30181 mesylate Type A showed 2.584% weight lossbefore 150° C., matching with a monohydrate weight loss (MW 802.849 forHM30181 mesylate salt monohydrate, 2.24% wt loss), followed by a 4.133%weight loss before 250° C., possibly due to disassociation anddecomposition (see FIG. 3 ). By ¹H-NMR, HM30181 mesylate Type A waspotentially a hydrate, as no solvents were detected other than water(see FIG. 4 ).

By DVS, HM30181 mesylate Type A was hygroscopic and absorbed 2.26% waterfrom 0-95% RH, with no change in XRPD pattern observed (see FIG. 5 andFIG. 6 ).

Cyclic DSC to 110° C. of HM30181 mesylate Type A resulted in no XRPDchange (see FIG. 7 and FIG. 8 ). Cyclic DSC of HM30181 mesylate Type Ato 185° C. resulted in HM30181 mesylate Type A with reducedcrystallinity (FIG. 9 and FIG. 10 ). Cyclic DSC of HM30181 mesylate TypeA to 200° C. resulted in amorphous material (see FIG. 11 and FIG. 12 ).Freebase HM3018-A (6004273-06-A) was also characterized by XRPD and wasfound to be amorphous (see FIG. 13 ). Based on ¹H-NMR data,decomposition of Type A initiated after heating to 200° C. (see FIG. 14).

Solubility of HM30181 mesylate starting material was estimated insolvents (see below), and results are listed in Table 2.

TABLE 2 Dissolved after Solubility heating to 50° C. Experiment IDSolvent (mg/mL) for 2 hours? B00505-05-C-1 MeOH 1.7-2.0 B00505-05-C-2EtOH <0.9 Almost B00505-05-C-3 IPA <1.0 No B00505-05-C-4 Acetone <1.0 NoB00505-05-C-5 MIBK <1.0 No B00505-05-C-6 EtOAc <0.9 No B00505-05-C-7IPOAc <1.0 No B00505-05-C-8 THF <1.0 No B00505-05-C-9 2-MeTHF <1.0 NoB00505-05-C-10 1,4-Dioxane <1.0 No B00505-05-C-11 MTBE <1.0 NoB00505-05-C-12 ACN <1.0 Yes B00505-05-C-13 DCM <1.0 Yes B00505-05-C-14CHCl₃ 5.0-6.7 B00505-05-C-15 Toluene <1.0 No B00505-05-C-16 n-Heptane<1.0 No B00505-05-C-17 H₂O <1.0 No B00505-05-C-18 Cyclohexane <1.0 NoB00505-05-C-19 MEK <1.0 No B00505-05-C-20 T-BuOH <1.0 No B00505-05-C-21NMP 4.8-6.3 B00505-05-C-22 DMSO 22-44 B00505-05-C-23 DMA 3.2-4.8B00505-05-C-24 n-Propanol <1.0 No B00505-05-C-25 n-Propyl acetate <1.0No B00505-05-C-26 Cumene <1.0 No

Slurry-based generation and/or screening for HM30181 mesylate polymorphswas performed by preparing slurries of starting material HM30181mesylate Type A in a variety of solvents and under a variety ofconditions as described below. The resulting solids were analyzed byXRPD and identified for physical state. Results are summarized in Table3 and Table 4.

Slurries of HM30181 mesylate Type A in MeOH generated HM30181 mesylateType B after 1 week at temperatures of 4° C. to 50° C. (see FIG. 15 ).Slurries of HM30181 mesylate Type A in DCM at ambient temperature andacetonitrile at 4° C. to 50° C. generated HM30181 mesylate Type C (seeFIG. 16 ). Slurries of HM30181 mesylate Type A in NMP at ambienttemperature generated HM30181 mesylate Type D (see FIG. 17 ). BothHM30181 mesylate Type C and HM30181 mesylate Type D showed significantsimilarities to HM30181 mesylate Type A. Slurries of HM30181 mesylateType A in DMA at ambient conditions generated HM30181 mesylate Type E(see FIG. 18 ). Slurries of HM30181 mesylate Type A in DMF generatedHM30181 mesylate Type F at ambient temperature (see FIG. 19 ) and Type Gat 50° C. (see FIG. 20 ).

Table 3 (continued in Table 4) API Solvent Crystal Exp Method Temp.Solvent (mg) (mL) Type 6007235-16-C10 Slurry 4° C. 1,4-Dioxane 25.9 0.2A 6007235-16-B10 Slurry 50° C. 1,4-Dioxane 23.4 0.2 A 6007235-16-A10Slurry ambient 1,4-Dioxane 21 0.2 A temperature 6007235-16-A34 Slurryambient 1-Methyl-2- 25 0.2 A temperature pyrrolidinone/water (1:1)6007235-16-C9 Slurry 4° C. 2-Methyl 27.4 0.2 A tetrahydrofuran6007235-16-B9 Slurry 50° C. 2-Methyl 24.5 0.2 A tetrahydrofuran6007235-16-A9 Slurry ambient 2-Methyl 28.7 0.2 A temperaturetetrahydrofuran 6007235-16-C4 Slurry 4° C. Acetone 25.3 0.2 A6007235-16-B4 Slurry 50° C. Acetone 27.9 0.2 A 6007235-16-A4 Slurryambient Acetone 26.7 0.2 A temperature 6007235-16-C13 Slurry 4° C. CHCl₃22.4 0.2 A 6007235-16-B13 Slurry 50° C. CHCl₃ 29 0.1 A 6007235-16-A14Slurry ambient CHCl₃ 25.6 0.2 A temperature 6007235-16-C21 Slurry 4° C.Cumene 22.1 0.2 A 6007235-16-B25 Slurry 50° C. Cumene 26.3 0.2 A6007235-16-A26 Slurry ambient Cumene 26.1 0.2 A temperature6007235-16-C17 Slurry 4° C. Cyclohexane 24.2 0.2 A 6007235-16-B17 Slurry50° C. Cyclohexane 23.8 0.2 A 6007235-16-A18 Slurry ambient Cyclohexane26.2 0.2 A temperature 6007235-16-C24 Slurry 4° C. Cyclopentylmethyl20.8 0.2 A ether 6007235-16-B29 Slurry 50° C. Cyclopentylmethyl 24.9 0.2A ether 6007235-16-A30 Slurry ambient Cyclopentylmethyl 21.7 0.2 Atemperature ether 6007235-16-A22 Slurry ambient DMSO 22.7 0.1 Atemperature 6007235-16-A31 Slurry ambient DMSO/water (1:1) 25.1 0.2 Atemperature 6007235-16-C2 Slurry 4° C. Ethanol 20.7 0.2 A 6007235-16-B2Slurry 50° C. Ethanol 26.9 0.2 A 6007235-16-A2 Slurry ambient Ethanol21.7 0.2 A temperature 6007235-16-C6 Slurry 4° C. Ethyl acetate 26.2 0.2A 6007235-16-B6 Slurry 50° C. Ethyl acetate 25.5 0.2 A 6007235-16-A6Slurry ambient Ethyl acetate 24.2 0.2 A temperature 6007235-16-C22Slurry 4° C. Ethyl formate 24.8 0.2 A 6007235-16-B27 Slurry 50° C. Ethylformate 25 0.2 A 6007235-16-A28 Slurry ambient Ethyl formate 25.1 0.2 Atemperature 6007235-16-C23 Slurry 4° C. Isobutyl Acetate 23.7 0.2 A6007235-16-B28 Slurry 50° C. Isobutyl Acetate 22 0.2 A 6007235-16-A29Slurry ambient Isobutyl Acetate 25.4 0.2 A temperature 6007235-16-C3Slurry 4° C. isopropanol 26.8 0.2 A 6007235-16-B3 Slurry 50° C.isopropanol 21.5 0.2 A 6007235-16-A3 Slurry ambient isopropanol 23.6 0.2A temperature 6007235-16-C7 Slurry 4° C. isopropyl acetate 25.9 0.2 A6007235-16-B7 Slurry 50° C. isopropyl acetate 21.9 0.2 A 6007235-16-A7Slurry ambient isopropyl acetate 28.8 0.2 A temperature 6007235-16-C18Slurry 4° C. methyl ethyl 21.8 0.2 A ketone 6007235-16-B18 Slurry 50° C.methyl ethyl 20.6 0.2 A ketone 6007235-16-A19 Slurry ambient methylethyl 27.1 0.2 A temperature ketone 6007235-16-C5 Slurry 4° C. methylisobutyl 26.1 0.2 A ketone 6007235-16-B5 Slurry 50° C. methyl isobutyl27 0.2 A ketone 6007235-16-A5 Slurry ambient methyl isobutyl 27.9 0.2 Atemperature ketone Table 4 (continued from Table 3) API Solvent CrystalExp Method Temp. Solvent (mg) (mL) Type 6007235-16-C11 Slurry 4° C.Methyl t-butyl ether 27.1 0.2 A 6007235-16-B11 Slurry 50° C. Methylt-butyl ether 27.5 0.2 A 6007235-16-A11 Slurry ambient Methyl t-butylether 26.8 0.2 A temperature 6007235-16-A32 Slurry ambient N,N- 26.9 0.2A temperature Dimethylacetamide/water (1:1) 6007235-16-A33 Slurryambient N,N- 27 0.2 A temperature Dimethylacetamide/water (1:1)6007235-16-C15 Slurry 4° C. n-Heptane 26.8 0.2 A 6007235-16-B15 Slurry50° C. n-Heptane 22.7 0.2 A 6007235-16-A16 Slurry ambient n-Heptane 22.80.2 A temperature 6007235-16-C19 Slurry 4° C. n-Propanol 22.3 0.2 A6007235-16-B23 Slurry 50° C. n-Propanol 21.8 0.2 A 6007235-16-A24 Slurryambient n-Propanol 23.3 0.2 A temperature 6007235-16-C20 Slurry 4° C.n-Propyl acetate 21.5 0.2 A 6007235-16-B24 Slurry 50° C. n-Propylacetate 27.4 0.2 A 6007235-16-A25 Slurry ambient n-Propyl acetate 26.50.2 A temperature 6007235-16-B19 Slurry 50° C. t-Butanol 23.7 0.2 A6007235-16-A20 Slurry ambient t-Butanol 23.9 0.2 A temperature6007235-16-C8 Slurry 4° C. Tetrahydrofuran 22.9 0.2 A 6007235-16-B8Slurry 50° C. Tetrahydrofuran 21.7 0.2 A 6007235-16-A8 Slurry ambientTetrahydrofuran 20.9 0.2 A temperature 6007235-16-C14 Slurry 4° C.Toluene 24.2 0.2 A 6007235-16-B14 Slurry 50° C. Toluene 25.7 0.2 A6007235-16-A15 Slurry ambient Toluene 24.9 0.2 A temperature6007235-16-C16 Slurry 4° C. Water 27.6 0.2 A 6007235-16-B16 Slurry 50°C. Water 27 0.2 A 6007235-16-C28 Slurry 4° C. 1-Methyl-2- 23.9 0.2amorphous + pyrrolidinonee/water A (1:1) 6007235-16-C26 Slurry 4° C.N,N- 26.4 0.2 amorphous + Dimethylacetamide/water A (1:1) 6007235-16-C27Slurry 4° C. N,N- 24 0.2 amorphous + Dimethylacetamide/water A (1:1)6007235-16-A17 Slurry ambient Water 22 0.2 amorphous + temperature A6007235-16-B20 Slurry 50° C.

23.7 0.1 amorphous 6007235-16-B21 Slurry 50° C. DMSO 23.8 0.1 amorphous6007235-16-C25 Slurry 4° C. DMSO/water (1:1) 24.3 0.2 amorphous6007235-16-A1 Slurry ambient Methanol 24.9 0.2 B temperature6007235-16-B1 Slurry 50° C. Methanol 27 0.1 B 6007235-16-C1 Slurry 4° C.Methanol 23.2 0.2 B 6007235-16-A12 Slurry ambient Acetonitrile 25.5 0.2C temperature 6007235-16-A13 Slurry ambient Dichloromethane 21.9 0.2 Ctemperature 6007235-16-B12 Slurry 50° C. Acetonitrile 22.7 0.1 C6007235-16-C12 Slurry 4° C. Acetonitrile 21.9 0.2 C 6007235-16-A21Slurry ambient 1-Methyl-2-pyrrolidinone 24.6 0.2 D temperature6007235-16-A23 Slurry ambient N,N-Dimethylacetamide 25.6 0.1 Etemperature 6007235-16-A27 Slurry ambient N,N-Dimethylacetamide 26.2 0.1F temperature 6007235-16-B26 Slurry 50° C. N,N-Dimethylacetamide 27.30.1 G

Generation and/or screening of HM30181 mesylate polymorphs was alsoperformed by preparing HM30181 mesylate Type A starting material forliquid and solid vapor diffusion as described below. Resulting solidswere analyzed by XRPD and identified for physical state. Results aresummarized in Table 5, Table 6, and Table 7. Liquid vapor diffusion ofMTBE into NMP solution yielded HM30181 mesylate Type D (see FIG. 21 ).Liquid vapor diffusion of MEK into NMP or DMA solution yielded HM30181mesylate Type D (see FIG. 21 ). Some loss of crystallinity was noted onair-dried HM30181 mesylate Type D, suggesting possible solvate. Liquidvapor diffusion of 2-MeTHF into DMA or DMF solution yielded HM30181mesylate Type E (see FIG. 22 ). Liquid vapor diffusion of MTBE into DMAsolution yielded HM30181 mesylate Type E, with a few additionaldiffraction peaks, which Inventors believe are attributable to HM30181mesylate Type I (FIGS. 4 to 8 ). Liquid vapor diffusion of ACN into DMSOsolution yielded HM30181 mesylate Type H (see FIG. 23 ). Liquid vapordiffusion of Acetone, Ethyl Acetate, or isopropyl acetate into DMAsolution yielded HM30181 mesylate Type I (see FIG. 24 ). Air-drying ofHM30181 mesylate Type I yielded HM30181 mesylate Type J (see FIG. 24 ).Liquid vapor diffusion of isopropyl acetate into DMSO solution followedby air drying also yielded HM30181 mesylate Type J.

TABLE 5 API Solvent Crystal Exp Temperature Solvent (mg) (mL) Type6007235-17-A1 ambient methanol 25.2 0.1 A temperature 6007235-17-A2ambient ethanol 22.1 0.1 A temperature 6007235-17-A3 ambient isopropanol21.4 0.1 A temperature 6007235-17-A4 ambient

24.8 0.1 A temperature 6007235-17-A5 ambient methyl isobutyl ketone 25.80.1 A temperature 6007235-17-A6 ambient ethyl acetate 22.9 0.1 Atemperature 6007235-17-A7 ambient isopropyl acetate 23.8 0.1 Atemperature 6007235-17-A8 ambient tetrahydrofuran 24.5 0.1 A temperature6007235-17-A9 ambient 2-methyl tetrahydrofuran 24.9 0.1 A temperature6007235-17-A10 ambient methyl t-butyl ether 27.1 0.1 A temperature6007235-17-A11 ambient acetonitrile 27.7 0.1 A temperature6007235-17-A12 ambient dichloromethane 24.9 0.1 A temperature6007235-17-A13 ambient CHCl₃ 29 0.1 A temperature 6007235-17-A14 ambientmethyl ethyl ketone 23.4 0.1 A temperature 6007235-17-A15 ambientt-butanol 28.2 0.1 A temperature 6007235-17-A16 ambient n-propanol 25.40.1 A temperature 6007235-17-A17 ambient ethyl formate 27.3 0.1 Atemperature 6007235-17-A18 ambient cyclopentylmethyl ether 25.1 0.1 Atemperature 6007235-17-A19 25° C. 25° C./60% RH 22.9 0.1 A6007235-17-A20 40° C. 40° C./75% RH 23.5 0.1 A 6007235-17-A21 40° C.100% RH(water) 22.7 0.1 A

Table 6 (continued on Table 7) Anti- API Solvent solvent Exp Temp.Solvent (mg) (mL) Anti-solvent (mL) Crystal Type 6007235-19-A1 ambientmethanol 22 16 ethyl formate 0.5 No solids temperature 6007235-19-A2ambient methanol 23 16 dichloromethane 0.5 No solids temperature6007235-19-B1 ambient DMSO 25 0.3 ethanol 2 No solids temperature6007235-19-B2 ambient DMSO 25 0.3 isopropanol 2 No solids temperature6007235-19-B3 ambient DMSO 25 0.3 acetone 2 Amorphous temperature6007235-19-B4 ambient DMSO 25 0.3 MIBK 2 Amorphous temperature6007235-19-B5 ambient DMSO 25 0.3 ethyl acetate 2 Amorphous temperature6007235-19-B7 ambient DMSO 25 0.3 tetrahydrofuran 2 Amorphoustemperature 6007235-19-B8 ambient DMSO 25 0.3 2-MeTHF 2 Amorphoustemperature 6007235-19-B9 ambient DMSO 25 0.3 methyl t-butyl 2 Amorphoustemperature ether 6007235-19-B11 ambient DMSO 25 0.3 dichloromethane 2Amorphous temperature and A 6007235-19-B12 ambient DMSO 25 0.3 methylethyl 2 Amorphous temperature ketone 6007235-19-B13 ambient DMSO 25 0.3t-butanol 2 No solids temperature 6007235-19-C1 ambient NMP 30 3 ethanol3 No solids temperature 6007235-19-C2 ambient NMP 30 3 isopropanol 3 Nosolids temperature 6007235-19-C3 ambient NMP 30 3 acetone 3 No solidstemperature 6007235-19-C4 ambient NMP 30 3 MIBK 3 No solids temperature6007235-19-C5 ambient NMP 30 3 ethyl acetate 3 Amorphous temperature andA 6007235-19-C6 ambient NMP 30 3 isopropyl acetate 3 No solidstemperature 6007235-19-C7 ambient NMP 30 3 tetrahydrofuran 3 No solidstemperature 6007235-19-C8 ambient NMP 30 3 2-MeTHF 3 No solidstemperature 6007235-19-C10 ambient NMP 30 3 acetonitrile 3 No solidstemperature 6007235-19-C11 ambient NMP 30 3 dichloromethane 3 No solidstemperature 6007235-19-C13 ambient NMP 30 3 t-butanol 3 No solidstemperature 6007235-19-D1 ambient DMF 30 3 ethanol 3 No solidstemperature 6007235-19-D2 ambient DMF 30 3 isopropanol 3 Amorphoustemperature 6007235-19-D4 ambient DMF 30 3 MIBK 3 No solids temperature6007235-19-D5 ambient DMF 30 3 ethyl acetate 3 Amorphous temperature6007235-19-D6 ambient DMF 30 3 isopropyl acetate 3 Amorphous temperature6007235-19-D7 ambient DMF 30 3 tetrahydrofuran 3 No solids temperature6007235-19-D9 ambient DMF 30 3 methyl t-butyl 3 Amorphous temperatureether 6007235-19-D10 ambient DMF 30 3 acetonitrile 3 No solidstemperature 6007235-19-D11 ambient DMF 30 3 dichloromethane 3 No solidstemperature 6007235-19-D12 ambient DMF 30 3 methyl ethyl 3 Amorphoustemperature ketone 6007235-19-D13 ambient DMF 30 3 t-butanol 3 No solidstemperature 6007235-19-E1 ambient DMA 20 4 ethanol 8 No solidstemperature 6007235-19-E2 ambient DMA 20 4 isopropanol 8 No solidstemperature 6007235-19-E4 ambient DMA 20 4 MIBK 8 No solids temperature6007235-19-E7 ambient DMA 20 4 tetrahydrofuran 8 No solids temperature6007235-19-E10 ambient DMA 20 4 acetonitrile 8 No solids temperature6007235-19-E11 ambient DMA 20 4 dichloromethane 8 No solids temperature6007235-19-E13 ambient DMA 20 4 t-butanol 8 No solids temperature Table7 (continued from Table 6) Anti- XRPD API Solvent solvent (wet XRPD Exp.Temp. (mg) Solvent (mL) Anti-solvent (mL) cake) (air-dry) 6007235-19-C12ambient 30 NMP 3 MEK 3 Type D — temperature 6007235-19-E12 ambient 20DMA 4 MEK 8 Type D Type D and temperature amorphous 6007235-19-C9ambient 30 NMP 3 MTBE 3 Type D — temperature 6007235-19-D8 ambient 30DMF 3 2-MeTHF 3 Type E E temperature 6007235-19-E8 ambient 20 DMA 42-MeTHF 8 Type E E temperature 6007235-19-E9 ambient 20 DMA 4 MTBE 8Type E E temperature and I 6007235-19-B10 ambient 25 DMSO 0.3 ACN 2 TypeH — temperature 6007235-19-E3 ambient 20 DMA 4 Acetone 8 Type I Type Jtemperature 6007235-19-E5 ambient 20 DMA 4 EtOAc 8 Type I Type Jtemperature 6007235-19-E6 ambient 20 DMA 4 iPrOAc 8 Type I Type Jtemperature 6007235-19-B6 ambient 25 DMSO 0.3 iPrOAc 3 — Type Jtemperature

Generation and/or screening of HM30181 mesylate polymorphs by coolingwas carried out by treating HM30181 mesylate Type A starting materialusing gradual or rapid (i.e. crash) cooling as described below. Theresulting solids were analyzed by XRPD and identified for physicalstate. Results are summarized in Table 8. Cooling experiments inacetonitrile and DCM yielded HM30181 mesylate Type C; no significantchange was noted on air-drying (see FIG. 25 ).

TABLE 8 Solvent/ Solvent/ Anti- Anti- API solvent XRPD XRPD Exp. Temp.Temp solvent (mg) (mL) (wet cake) (air-dry) 6007235-17-C6 Slow Cooling50° C.-->4° C.  NMP 20.9 4 No solids 6007235-17-C14 Crash Cooling 50°C.-->−20° C. NMP 26.2 4 Amorphous 6007235-17-C2 Slow Cooling 50° C.-->4°C.  CHCl₃ 25.4 4 Amorphous and A 6007235-17-C10 Crash Cooling 50°C.-->−20° C. CHCl₃ 25.4 4 No solids 6007235-17-C13 Crash Cooling 50°C.-->−20° C. DCM 22.1 10 No solids 6007235-17-C3 Slow Cooling 50°C.-->4° C.  EtOH 5 20 No solids 6007235-17-C11 Crash Cooling 50°C.-->−20° C. EtOH 5 20 No solids 6007235-17-C9 Crash Cooling 50°C.-->−20° C. MeOH 22.9 10 No solids 6007235-17-C1 Slow Cooling 50°C.-->4° C.  MeOH 24.7 20 No solids 6007235-17-C7 Slow Cooling 50°C.-->4° C.  DMA 25 4 Amorphous and A 6007235-17-C15 Crash Cooling 50°C.-->−20° C. DMA 26.9 4 Amorphous 6007235-17-C8 Slow Cooling 50° C.-->4°C.  DMF 25.1 4 No solids 6007235-17-C16 Crash Cooling 50° C.-->−20° C.DMF 25.1 4 Amorphous 6007235-17-C4 Slow Cooling 50° C.-->4° C.  ACN 12.610 Type C Type C 6007235-17-C5 Slow Cooling 50° C.-->4° C.  DCM 26.8 10Type C + Type C peaks 6007235-17-C12 Crash cooling 50° C.-->−20° C. ACN12.6 10 Type C Type C

HM30181 mesylate was also subjected to evaporation methods by treatingHM30181 mesylate Type A starting material as described below. Theresulting solids were analyzed by XRPD and identified for physicalstate. Results are shown in Table 9.

TABLE 9 API Solvent Exp Temp. Solvent (mg) (mL) Crystal Type6007235-17-B6 ambient temperature methanol 12 10 Amorphous 6007235-17-B1ambient temperature methanol 12 10 Amorphous 6007235-17-B8 50° C.ethanol 5 20 Amorphous and A 6007235-17-B3 50° C. ethanol 5 20 Amorphous6007235-17-B5 50° C. dichloromethane 21.2 20 Amorphous 6007235-17-B1050° C. dichloromethane 28 20 Amorphous 6007235-17-B7 ambient temperatureCHCl₃ 27.4 7 Amorphous 6007235-17-B2 ambient temperature CHCl₃ 26 7Amorphous 6007235-17-B9 50° C. acetonitrile 10.5 40 Amorphous6007235-17-B4 50° C. acetonitrile 10.5 40 Amorphous

Generation and/or screening of HM30181 mesylate polymorphs by treatmentwith anti-solvents was performed by treating HM30181 mesylate Type Astarting material as described below. The resulting solids were analyzedby XRPD and identified for physical state. Results are summarized inTable 10, Table 11, Table 12, and Table 13. Anti-solvent studies in DMSOyielded HM30181 mesylate Type C (see FIG. 26 ). Anti-solvent studies inN,N-dimethylacetamide with methyl t-butyl ether yielded a HM30181mesylate Type F polymorphism (see FIG. 27 ). Anti-solvent addition andreverse anti-solvent addition in DMSO/EtOAc, DMA/MIBK, DMA/toluene andNMP/t-BuOH yielded primarily amorphous content and a polymorphism withsome similarity to HM30181 mesylate Type J (see FIG. 28 ). Otheranti-solvent studies in DMSO and DMF yielded a HM30181 mesylate Type Kpolymorphism (or possibly a mixture of types, see FIG. 29 ).Anti-solvent experiments in DMF/n-propanol and DMA/isopropanol yielded aHM30181 mesylate Type L polymorphism (see FIG. 30 ). Anti-solventaddition DMF/Toluene and DMA/t-BuOH were mostly amorphous, but alsogenerated HM30181 mesylate Type M (see FIG. 31 ).

Table 10 (continued on Table 11) Anti- Anti- solvent Evap NB Solventsolvent (mL) Precipitation? solids? Identification 6004273-06-A1 DMSOEtOH 20 N Y Type K 6004273-06-A2 DMSO IPA 10 Y insufficient solids/gel6004273-06-A3 DMSO Acetone 10 Y Type K 6004273-06-A4 DMSO MIBK 10 Y TypeK 6004273-06-A5 DMSO EtOAc 10 Y insufficient solids/gel 6004273-06-A6DMSO iPrOAc 10 Y insufficient solids/gel 6004273-06-A7 DMSO THF 10 YSimilar to Type K 6004273-06-A8 DMSO 2-MeTHF 10 Y Amorphous6004273-06-A9 DMSO 1,4-Dioxane 20 Y insufficient solids/gel6004273-06-A10 DMSO MTBE 20 Y Amorphous 6004273-06-A11 DMSO ACN 20 N YType C 6004273-06-A12 DMSO DCM 20 N Y Similar to Type C 6004273-06-A13DMSO CHCl₃ 20 N Y Type K 6004273-06-A14 DMSO Toluene 10 Y Amorphous6004273-06-A15 DMSO Water 20 N N insufficient solids/gel 6004273-06-A16DMSO MEK 10 Y insufficient solids/gel 6004273-06-A17 DMSO t-Butanol 10 YType K 6004273-06-A18 DMSO n-Propanol 10 Y Type K 6004273-06-A19 DMSOn-Propyl 10 Y Type K acetate 6004273-06-A20 CHCl₃ Ethanol 10 Y Type A6004273-06-A21 CHCl₃ IPA 10 Y Type A 6004273-06-A22 CHCl₃ Acetone 10 YType A 6004273-06-A23 CHCl₃ MIBK 10 Y Type A 6004273-06-A24 CHCl₃ EtOAc10 Y Type A 6004273-06-A25 CHCl₃ iPrOAc 10 Y Type A 6004273-06-A26 CHCl₃THF 10 Y Type A 6004273-06-A27 CHCl₃ 2-MeTHF 10 Y Type A 6004273-06-A28CHCl₃ 1,4-Dioxane 10 Y insufficient solids/gel 6004273-06-A29 CHCl₃ MTBE10 Y Type A 6004273-06-A30 CHCl₃ ACN 10 Y Type A 6004273-06-A31 CHCl₃DCM 10 Y Type A 6004273-06-A32 CHCl₃ Toluene 10 Y Type A 6004273-06-A33CHCl₃ n-Heptane 10 Y Type A Table 11 (continued from Table 10 and onTable 12) Anti- solvent Evap NB Solvent Anti-solvent (mL) Precipitation?solids? Identification 6004273-06-A34 CHCl₃ MeOAc 10 Y Type A6004273-06-A35 CHCl₃ Cyclohexane 10 Y Type A 6004273-06-A36 CHCl₃ MEK 10Y Type A 6004273-06-A37 CHCl₃ t-Butanol 10 Y Type A 6004273-06-A38 CHCl₃n-Propanol 10 Y Type A 6004273-06-A39 CHCl₃ n-Propyl 10 Y Type A acetate6004273-06-A40 CHCl₃ Ethyl formate 10 Y Type A 6004273-06-A41 CHCl₃iBuOAc 10 Y Type A 6004273-06-A42 CHCl₃ CPME 10 Y Type A 6004273-06-A43DMF EtOH 20 N Y Amorphous 6004273-06-A44 DMF IPA 10 Y Type K6004273-06-A45 DMF Acetone 20 N Y insufficient solids/gel 6004273-06-A46DMF MIBK 10 Y Amorphous 6004273-06-A47 DMF EtOAc 10 Y Type A6004273-06-A48 DMF iPrOAc 10 Y insufficient solids/gel 6004273-06-A49DMF THF 20 N Y insufficient solids/gel 6004273-06-A50 DMF 2-MeTHF 10 Yinsufficient solids/gel 6004273-06-A51 DMF 1,4-Dioxane 20 N Yinsufficient solids/gel 6004273-06-A52 DMF MTBE 10 Y Type A6004273-06-A53 DMF ACN 20 N Y insufficient solids/gel 6004273-06-A54 DMFDCM 20 N Y insufficient solids/gel 6004273-06-A55 DMF CHCl₃ 20 N Yinsufficient solids/gel 6004273-06-A56 DMF Toluene 10 Y Type M6004273-06-A57 DMF Water 20 N Y insufficient solids/gel 6004273-06-A58DMF MEK 20 N Y insufficient solids/gel 6004273-06-A59 DMF t-Butanol 10 Yinsufficient solids/gel 6004273-06-A60 DMF n-Propanol 20 N Y Type L +amorphous 6004273-06-A61 DMF n-Propyl 10 Y Type A acetate 6004273-06-A62NMP EtOH 20 N N insufficient solids/gel 6004273-06-A63 NMP IPA 10 Yinsufficient solids/gel 6004273-06-A64 NMP Acetone 20 N N insufficientsolids/gel 6004273-06-A65 NMP MIBK 10 Y insufficient solids/gel6004273-06-A66 NMP EtOAc 10 Y insufficient solids/gel Table 12(continued from Table 11) Anti- solvent Evap NB Solvent Anti-solvent(mL) Precipitation? solids? Identification 6004273-06-A67 NMP iPrOAc 10Y insufficient solids/gel 6004273-06-A68 NMP THF 20 N N insufficientsolids/gel 6004273-06-A69 NMP 2-MeTHF 10 Y insufficient solids/gel6004273-06-A70 NMP 1,4-Dioxane 20 N N Type A 6004273-06-A71 NMP MTBE 10Y insufficient solids/gel 6004273-06-A72 NMP ACN 20 N N insufficientsolids/gel 6004273-06-A73 NMP DCM 20 N N insufficient solids/gel6004273-06-A74 NMP CHCl₃ 20 N N insufficient solids/gel 6004273-06-A75NMP Toluene 10 Y insufficient solids/gel 6004273-06-A76 NMP Water 20 N Ninsufficient solids/gel 6004273-06-A77 NMP MEK 20 N Y Amorphous6004273-06-A78 NMP t-BuOH 20 hazy Y Amorphous + Type J 6004273-06-A79NMP n-Propanol 20 N N insufficient solids/gel 6004273-06-A80 NMPn-Propyl 10 Y insufficient acetate solids/gel 6004273-06-A81 DMA EtOH 20N N insufficient solids/gel 6004273-06-A82 DMA IPA 20 hazy Y Type L +amorphous 6004273-06-A83 DMA Acetone 20 N Y insufficient solids/gel6004273-06-A84 DMA MIBK 10 Y Amorphous + Type J 6004273-06-A85 DMA EtOAc10 Y insufficient solids/gel 6004273-06-A86 DMA iPrOAc 10 Y insufficientsolids/gel 6004273-06-A87 DMA THF 20 N N insufficient solids/gel6004273-06-A88 DMA 2-MeTHF 10 Y insufficient solids/gel 6004273-06-A89DMA 1,4-Dioxane 20 N N insufficient solids/gel 6004273-06-A90 DMA MTBE10 Y Amorphous + Type F 6004273-06-A91 DMA ACN 20 N N insufficientsolids/gel 6004273-06-A92 DMA DCM 20 N N insufficient solids/gel6004273-06-A93 DMA CHCl₃ 20 N N insufficient solids/gel 6004273-06-A94DMA Toluene 10 Y Amorphous + Type J 6004273-06-A95 DMA Water 20 N Yinsufficient solids/gel 6004273-06-A96 DMA MEK 20 N N insufficientsolids/gel 6004273-06-A97 DMA t-Butanol 20 N Y Type M 6004273-06-A98 DMAn-Propanol 20 N Y insufficient solids/gel 6004273-06-A99 DMA n-Propyl 10Y insufficient acetate solids/gel

TABLE 13 Anti- solvent Evap NB Solvent Anti-solvent (mL) Precipitation?solids? Identification 6004273-06-A100 NMP 1,4-Dioxane 20 N insufficientsolids/gel 6004273-06-A101 NMP MIBK 10 Y insufficient solids/gel6004273-06-A102 CHCl3 EtOH 10 Y Type A 6004273-06-A103 CHCl3 MTBE 10 YType A 6004273-06-A104 CHCl3 n-Heptane 10 Y Type A 6004273-06-A105 DMSOEtOAc 10 Y insufficient solids/gel 6004273-06-A106 DMSO Toluene 10 YAmorphous 6004273-06-A107 DMSO Water 20 N insufficient solids/gel6004273-06-A108 DMA Acetone 20 N insufficient solids/gel 6004273-06-A109DMA iPrOAc 10 Y insufficient solids/gel 6004273-06-A110 DMF IPA 10 YAmorphous + Type K 6004273-06-A111 DMF THF 20 N insufficient solids/gel

Large scale studies were performed with HM30181 mesylate Type A startingmaterial on a 200 mg scale as described below. Results are summarized inTable 14. Solids were isolated by vacuum filtration. The wet cakes fromfiltration from DMA, DMF, and NMP slurries were washed with 1 mL to 2 mLmethanol to remove solvents. Solids were then vacuum dried at 80° C.overnight.

At large scale, a slurry of HM30181 mesylate Type A starting material inmethanol at ambient conditions generated a mixture of HM30181 mesylateType B and HM30181 mesylate Type A after 9 days (see FIG. 32 ). After 14days, the slurry in methanol generated a HM30181 mesylate Type Npolymorph. HM30181 mesylate Type N showed some loss of crystallinityafter vacuum-drying, suggesting a methanol solvate (see FIG. 32 ). Aslurry of HM30181 mesylate Type A starting material in acetonitrile atambient conditions generated HM30181 mesylate Type C after 14 days; noloss of crystallinity was detected after vacuum-drying (see FIG. 33 ). Ascaled-up slurry of HM30181 mesylate Type A starting material in NMPprovided a primarily amorphous material (see FIG. 34 ). A scaled-upslurry of HM30181 mesylate Type A starting material in DMA yieldedHM30181 mesylate Type E after 6 days (see FIG. 35 ). HM30181 mesylateType E showed no loss of crystallinity after vacuum drying. A scaled-upslurry of HM30181 mesylate Type A starting material in DMF at 50° C.yielded HM30181 mesylate Type F rather than the expected HM30181mesylate Type G after 9 days (see FIG. 36 ). Some change of pattern wasnoted after vacuum-drying. A scaled-up slurry of HM30181 mesylate Type Astarting material in DMF at ambient temperature initially showed nochange from Type A (see FIG. 37 ). This slurry was heated in an attemptto generate HM30181 mesylate Type G. A scaled-up slurry of HM30181mesylate Type A starting material in DMF at 100° C. yielded HM30181mesylate Type F after 2 days (see FIG. 37 ). A scaled-up slurry ofHM30181 mesylate Type A starting material in DMF at 150° C. yielded apreviously unobserved XRPD pattern after 5 days (see FIG. 37 ). ¹H-NMRresults in DMSO-d₆ suggested that the DMF slurry at 150° C. resulted indegradation, as the ¹H-NMR spectrum did not match either the startingmaterial or freebase (see FIG. 38 ).

By ¹H-NMR, no disproportionation or degradation was detected in HM30181mesylate Types C, E, F and N polymorphs (see FIG. 39 ). By ¹H-NMR,HM30181 mesylate Type E contained DMA, HM30181 mesylate Type F containsDMF, and HM30181 mesylate Type N contains MeOH.

TABLE 14 Anti- Solvent Conc. Anti- solvent Desired NB Method Solvent(mL) (mg/mL) solvent (mL) Type Observations 6004273-10-B Slurry at MeOH0.2 480 B Type N ambient temperature 6004273-10-C Slurry at MeCN 0.2 545C Type C ambient temperature 6004273-10-D Slurry at NMP 0.2 550 DAmorphous ambient temperature 6004273-10-E Slurry at DMA 0.2 576 E TypeE ambient temperature 6004273-10-F Slurry at DMF 0.2 595 F Type Aambient temperature 6004273-10- Slurry at DMF 0.2 595 G Type F F-100C100° C. (for 2 days) 6004273-10- Slurry at DMF 0.2 595 G DecompositionF-150C 150° C. (for 5 d) 6004273-10-G Slurry at DMF 0.2 576 G Type F 50°C. 6004273-10-H Liquid DMSO 5 60 MeCN 3 H No precipitation, vaporevaporated- no sorption solids due to DMSO 6004273-10-J Liquid DMSO 1060 Acetone 3 J No precipitation, vapor evaporated- no sorption solidsdue to DMSO 6004273-10-K Anti- DMSO 10 60 MIBK 200 K Amorphous solventaddition 6004273-10-L Anti- DMF 20 5 n-propanol 200 L Amorphous solventaddition 6004273-10-M Anti- DMA 20 5 t-BuOH 200 M Amorphous solventaddition 6004273-10-M1 Anti- DMF 20 5 Toluene 300 M Amorphous solventaddition

Characteristics of HM30181 mesylate salt polymorphisms as characterizedby XRPD, TGA, DSC, and DVS are summarized below:

-   -   HM30181 mesylate Type A represents a prior art preparation of        HM30181 mesylate that can be used as a starting material in        generation of novel polymorphs of this compound.    -   Crystalline HM30181 mesylate Type B was obtained through        slurrying in methanol at 4° C. to 50° C. and is distinct by XRPD        (see FIG. 40 ). By DSC, HM30181 mesylate Type B displayed an        endotherm at 159.92° C. (see FIG. 41 ). By TGA, HM30181 mesylate        Type B showed 1.865% weight loss before 170° C., followed by        possible disassociation and decomposition (see FIG. 41 ).    -   Crystalline HM30181 mesylate Type C polymorph was obtained        through slurrying in acetonitrile at ambient temperature and        shows characteristic results on XRPD. No change in crystalline        pattern was noted after vacuum drying overnight (see FIG. 42 ).        By DSC, HM30181 mesylate Type C displays an endotherm at        159.60° C. (see FIG. 43 ). By TGA, HM30181 mesylate Type C        showed 2.987% weight loss before 170° C., followed by a        disassociation and decomposition (see FIG. 43 ). By 1H-NMR,        HM30181 mesylate Type C is confirmed to contain only water and        no acetonitrile, (˜2.07 ppm) suggesting a monohydrate (see FIG.        44 ).    -   Crystalline HM30181 mesylate Type D was obtained through        slurrying in N-methyl pyrrolidone at ambient temperature and        shows characteristic results by XRPD. No change in crystalline        pattern was noted after air drying overnight (see FIG. 45 ). By        DSC/TGA, HM30181 mesylate Type D displayed an endotherm at        66.97° C. and a 14.71% weight loss before 150° C., suggesting        significant residual solvent content (see FIG. 46 ).    -   Crystalline HM30181 mesylate Type E was obtained through        slurrying in N,N-dimethylacetamide at ambient temperature and        shows characteristic results by XRPD. No change in crystalline        pattern was noted after vacuum drying overnight at 80° C. (see        FIG. 47 ). By DSC/TGA, HM30181 mesylate Type E displayed an        endotherm at 154.42° C. and a 5.247% weight loss before 168° C.,        suggesting HM30181 mesylate Type E was a solvate (see FIG. 48 ).        By ¹H-NMR, HM30181 mesylate Type E was confirmed to contain DMA        (see FIG. 49 ).    -   Crystalline HM30181 mesylate Type F was obtained through        slurrying in dimethylformamide at 50° C. and shows        characteristic results by XRPD (see FIG. 50 ). A significant        reduction in crystallinity was noted after vacuum drying        overnight at 80° C., suggesting HM30181 mesylate Type F is a        metastable solvate. By DSC/TGA, HM30181 mesylate Type F        displayed an endotherm at 148.41° C. and a 5.123% weight loss        before 180° C., consistent with loss of DMF (see FIG. 51 ). By        ¹H-NMR, HM30181 mesylate Type F was confirmed to contain DMF        (see FIG. 52 ).    -   Crystalline HM30181 mesylate Type G was obtained through        slurrying in dimethylformamide at 50° C. and shows        characteristic results by XRPD. No reduction in crystallinity        was noted after air drying overnight (see FIG. 53 ). By DSC/TGA,        HM30181 mesylate Type G displayed an endotherm at 69.02° C. and        at 233.29° C. and a 1.451% weight loss before 150° C., which was        consistent with loss of DMF (see FIG. 54 ).    -   Crystalline HM30181 mesylate Type H was obtained through liquid        vapor diffusion of acetonitrile into a DMSO stock of HM30181        mesylate and shows characteristic results by XRPD (see FIG. 55        ). By DSC/TGA, HM30181 mesylate Type H displayed an endotherm at        126.52° C. and a 7.444% weight loss before 200° C., potentially        from residual ACN and DMSO or from a solvate (see FIG. 56 ).    -   Crystalline HM30181 mesylate Type I was obtained through liquid        vapor diffusion of acetone, ethyl acetate, or isopropyl acetate        into a DMA stock of HM30181 mesylate and shows characteristic        results by XRPD (see FIG. 57 ). Air-drying of HM30181 mesylate        Type I yielded HM30181 mesylate Type J.    -   HM30181 mesylate Type J was obtained through liquid vapor        diffusion of acetone, ethyl acetate, or isopropyl acetate into a        DMA stock or isopropyl acetate into a DMSO stock of HM30181        mesylate followed by air-drying. By XRPD, HM30181 mesylate Type        J is crystalline (see FIG. 58 ). By TGA, HM30181 mesylate Type J        displayed a 2.887% weight loss before 150° C., suggesting a        solvate or hydrate (see FIG. 59 ).    -   HM30181 mesylate type K was obtained through anti-solvent        addition using DMF/IPA and multiple DMSO systems (ethanol,        acetone, MIBK, THF, chloroform, t-butanol, n-propyl acetate, and        n-propanol). By XRPD, HM30181 mesylate Type K is partially        crystalline (see FIG. 60 ).    -   HM30181 mesylate Type L was obtained through anti-solvent        addition in DMF/n-propanol and DMA/isopropanol systems. By XRPD,        HM30181 mesylate Type L is partially crystalline (see FIG. 61 ).    -   HM30181 mesylate type M was obtained through anti-solvent        addition in DMF/toluene and DMA/t-butanol systems. By XRPD,        HM30181 mesylate Type M is partially crystalline (see FIG. 62 ).

Crystalline HM30181 mesylate Type N was obtained after 14-days treatmentof HM30181 mesylate Type A starting material as a slurry in methanol atambient temperature (see FIG. 63 ). HM30181 mesylate Type N showed someloss of crystallinity after vacuum-drying, suggesting a methanolsolvate. By DSC, HM30181 mesylate Type N displayed endotherms at 159.28°C. and 188.47° C. with a 2.128% weight loss before 180° C., followed bypossible disassociation and decomposition (see FIG. 64 ). ¹H-NMRconfirmed the presence of methanol (see FIG. 65 ).

HM30181 mesylate Type C and E forms were further analyzed to determineunit cell dimensions. Unit cell parameters for the Type C polymorph ofHM30181 mesylate were calculated using cumulative XRPD spectra, peakidentifications for which are shown in Table 15. Notably distinct peaksfor HM30181 mesylate Type C are shown in bold and italicized in Table15. Estimated values of unit cell parameters derived from the Type Cpolymorph of HM30181 mesylate are shown in Table 16 and are consistentwith triclinic P unit cells.

TABLE 15 FWHM Rel. Pos. Height Left d-spacing Int. [°2Th.] [cts] [°2Th.][Å] [%] 3.5 34.6 0.05 25.5 1.8 5.1 95.5 0.115 17.5 5.0 5.6 30.2 0.0815.8 1.6

9.9 37.7 0.20 8.9 2.0 11.7 305.9 0.20 7.6 16.2 12.1 322.7 0.10 7.3 17.112.8 1892.0 0.10 6.9 100.0 13.9 205.8 0.13 6.4 10.9 15.0 609.3 0.13 5.932.2 15.5 113.8 0.82 5.7 6.0 16.1 253.9 0.15 5.5 13.4 17.8 71.2 0.46 5.03.8 19.2 439.5 0.20 4.6 23.2 19.7 590.3 0.20 4.5 31.2 20.0 362.7 0.204.4 19.2 21.2 60.7 0.10 4.2 3.2 22.1 20.0 0.10 4.0 1.0 22.9 309.0 0.133.9 16.3 23.4 99.7 0.18 3.8 5.3 24.5 37.2 0.20 3.6 2.0 25.1 63.6 0.313.6 3.4 26.1 1066.2 0.31 3.4 56.4 26.9 130.0 0.20 3.3 6.9 28.1 144.20.23 3.2 7.6 29.4 15.4 0.31 3.0 0.8 32.4 19.0 0.20 2.8 1.0 34.3 25.90.05 2.6 1.4 35.50 13.2 0.41 2.5 0.7 36.5 16.1 0.15 2.5 0.9 37.3 12.00.08 2.4 0.6

TABLE 16 Reflection Conditions Crystal System Triclinic Bravais TypePrimitive (P) Space Group Instrument Settings Goniometer Radius (mm)240.00 Unit Cell a (Å) 7.34 (2) b (Å) 14.6 (1) c (Å) 17.5 (8) Alpha (°)51.8 (6) Beta (°) 62.3 (2) Gamma (°) 90.4 (3) Volume (Å³) 1179.70Refinement Results No. Unindexed Lines 0 No. Indexed Lines 17 Total No.5110 Calculated Lines Chi Square 3.777537E−0006 Snyder's FOM 2.0791

Characteristic XRPD peak values for the Type E polymorph are provided inTable 17, where notably distinct peaks are indicated by bolded anditalicized numerals. It should be appreciated that these are distinctand different from those of polymorph Type C, indicating that the Type Cand Type E polymorphs are distinct and different from one another andthat both Type C and Type E polymorphs are distinct and different fromthe prior art Type A polymorph of HM30181 mesylate.

TABLE 17 FWHM Rel. Pos. Height Left d-spacing Int. [°2Th.] [cts] [°2Th.][Å] [%]

5.2 4.1 0.05 17.0 0.4 6.0 16.0 0.038 14.7 1.5 6.2 13.5 0.051 14.3 1.36.3 22.1 0.038 14.0 2.1 8.2 1047.3 0.15 10.8 100.0 9.0 35.8 0.038 9.93.4 9.1 29.7 0.15 9.7 2.8 9.9 13.9 0.05 9.0 1.3

11.6 136.8 0.20 7.6 13.1 12.3 114.7 0.18 7.2 11.0 12.9 170.3 0.15 6.916.3 13.6 30.8 0.15 6.5 2.9

15.4 227.7 0.13 5.7 21.8 15.9 801.8 0.23 5.6 76.6 16.3 46.5 0.15 5.4 4.4

17.7 281.7 0.20 5.0 26.9 18.3 200.1 0.23 4.9 19.1 18.9 298.0 0.20 4.728.4 19.3 164.2 0.13 4.6 15.7 19.6 106.5 0.13 4.5 10.2 20.0 103.1 0.154.4 9.8 20.4 153.2 0.13 4.3 14.6

21.3 320.9 0.13 4.2 30.6 21.6 243.7 0.10 4.1 23.3 22.0 111.1 0.20 4.010.6 22.4 128.7 0.15 4.0 12.3 22.7 418.2 0.26 3.9 39.9

24.1 116.9 0.10 3.7 11.2 24.6 174.5 0.20 3.6 16.7 24.9 214.9 0.18 3.620.5 26.2 994.9 0.15 3.4 95.0

 

28.3 94.3 0.18 3.2 9.0 28.8 20.2 0.15 3.1 1.9 29.3 62.1 0.15 3.0 5.930.7 45.9 0.20 2.9 4.4 32.1 117.5 0.18 2.8 11.2 33.3 40.2 0.15 2.7 3.833.9 42.7 0.31 2.6 4.1 34.7 33.4 0.26 2.6 3.2 35.7 22.6 0.20 2.5 2.237.0 29.8 0.15 2.4 2.8Unit cell parameters for the Type E polymorph of HM30181 mesylate werecalculated using cumulative XRPD spectra. Estimated values of unit cellparameters derived from the Type E polymorph of HM30181 mesylate areshown in Table 18 and are consistent with triclinic P unit cells.

TABLE 18 Reflection Conditions Crystal System Triclinic Bravais TypePrimitive (P) Space Group Instrument Settings Goniometer Radius (mm) 240.00 Unit Cell a (Å) 8.2 (2) b (Å) 9.8 (2) c (Å) 23.7 (4) Alpha (°)75.2 (2) Beta (°) 78.66 (3) Gamma (°) 111.69 (3) Volume (Å³) 1618.45Refinement Results No. Unindexed Lines  0 No. Indexed Lines 32  TotalNo. 6088   Calculated Lines Chi Square 8.802423E−0007 Snyder's FOM   5.7186

As noted above, HM30181 is an inhibitor of P-glycoprotein, an effluxtransport protein that is effective at removing a wide range oftherapeutic from cells and forms an important part of the blood brainbarrier. While this function is essentially protective, it can adverselyimpact the use of therapeutic drugs that P-glycoprotein substrates.Examples of drugs that are transported by P-glycoprotein include, butare not limited to, antineoplastic drugs (e.g., docetaxel, etoposide,vincristine), calcium channel blockers (e.g., amlodipine), calcineurininhibitors (e.g., cyclosporin, tacrolimus), digoxin, macrolideantibiotics (e.g., clarithromycin), and protease inhibitors.Accordingly, HM30181 mesylate can be used to alter the pharmacokineticsof therapeutic drug substrates of P-glycoprotein by reducing efflux ofsuch drugs from the cells of an individual undergoing treatment.

Conventional process for the production of HM30181 provide the Type Apolymorph. Inventors have produced and identified a number of otherforms of this compound, including Type B, Type C, Type D, Type E, TypeF, Type G, Type H, Type I, Type J, Type K, Type L, Type M, and Type Npolymorphs of HM30181. As shown above, these are different and distinctfrom the prior art Type A polymorph and from each other. Inventorsbelieve that these new polymorphs of HM30181 can provide differentstabilities and/or pharmacokinetics (e.g., rate of absorption, etc.)than those provided by the prior art Type A polymorph.

Accordingly, another embodiment of the inventive concept is theapplication of one or more of a Type B, Type C, Type D, Type E, Type F,Type G, Type H, Type I, Type J, Type K, Type L, Type M, and/or Type Npolymorph of HM30181 to inhibit P-glycoprotein, and in turn alter thepharmacokinetics of a drug that is a substrate of P-glycoprotein. Insome of such embodiments the drug can be a chemotherapeutic drug used inthe treatment of cancer.

In such embodiments one or more of a Type B, Type C, Type D, Type E,Type F, Type G, Type H, Type I, Type J, Type K, Type L, Type M, and/orType N polymorph of HM30181 can be administered in concert with a drugthat is a P-glycoprotein substrate to an individual that is in need oftreatment for a disease or condition that is responsive to such a drug.In some embodiments a Type B, Type C, Type D, Type E, Type F, Type G,Type H, Type I, Type J, Type K, Type L, Type M, and/or Type N polymorphof HM30181 can be provided as a separate formulation. Alternatively, oneor more of a Type B, Type C, Type D, Type E, Type F, Type G, Type H,Type I, Type J, Type K, Type L, Type M, and/or Type N polymorph ofHM30181 can be formulated in combination with a drug that is aP-glycoprotein substrate. In a preferred embodiment the disease iscancer, and the drug that is a P-glycoprotein substrate is achemotherapeutic drug used to treat cancer.

Methods

As noted above, polymorphs of HM30181 mesylate were provided bytreatment of a conventional HM30181 mesylate Type A preparation with avariety of solvents, and using a range of techniques. For solubilitystudies of HM30181 mesylate Type A in a variety of solvents a sample (˜2mg) of the solid was transferred into a 4-mL glass vial. Solvent wasadded to the vial in a stepwise fashion, 50 μL per step until 100 μLtotal volume followed by 100 μL per step until concentration was lessthan 1.0 mg/mL. Samples were mixed thoroughly after each addition bysonication for 2 minutes and vortexing for 1 minute. Volumes of solvent(V1 and V2) were recorded and used to estimate solubility. Solvents usedare summarized below in Table 19.

TABLE 19 Abbreviation Solvent Abbreviation Solvent MeOH Methanol THFTetrahydrofuran EtOH Ethanol 2-MeTHF 2-Methyltetrahydrofuran IPAIsopropyl DMF Dimethyl formamide alcohol ACN Acetonitrile DMSO Dimethylsulfoxide MIBK Methyl isobutyl CHCl₃ Chloroform ketone EtOAc Ethylacetate DCM Dichloromethane iPrOAc Isopropyl DMAc Dimethylacetamideacetate MTBE Methyl tert-butyl t-BuOH t-Butanol ether

Screening of polymorphisms of HM30181 mesylate can include preparationof a slurry. Typically, a slurry was prepared by suspending 5 mg to 20mg of sample in 0.1 mL to 0.5 mL solvent in a 1.5 mL or 3.0 mL glassvial. The suspension a was stirred at target temperature (e.g. 4° C.,ambient temperature, 50° C.) at 200 rpm. Solids for X-ray powderdiffraction (XRPD) analysis were separated by centrifuging at 14,000 rpmfor 5 minutes at ambient temperature. If no solid or gel is obtained,the slurry can be move to a fume hood for evaporation of the solvent.

In some embodiments anti-solvent addition was used. In this method aconcentrated stock of compound in solvent is provided and ananti-solvent quickly added to the concentrated solution while stirringto induce precipitation. Solids can be isolated for XRPD analysis usingfiltration or centrifugation.

In some embodiments reverse anti-solvent addition was used. In thismethod a concentrated stock of compound in solvent is provided andquickly added to an anti-solvent with stirring to induce precipitation.Solids can be isolated for XRPD analysis using filtration orcentrifugation.

In some embodiments slow cooling was used. In this method a concentratedsuspension of compound in solvent is provided. This solution was heatedto 50° C. and held at 50° C. for at least 30 minutes. The resultingsolution or suspension was filtered at 50° C. using a 0.45 micron PTFEfilter and the filtrate collected into clean vials. The resulting clearsolution was cooled to 5° C. to induce precipitation. Solids wereisolated solids for XRPD analysis using filtration or centrifugation.

In some embodiments crash cooling was used. In this method aconcentrated suspension of compound in solvent is provided. Thesuspension was heated to 50° C. and held at 50° C. for at least 30minutes. The heated solution or suspension was filtered at 50° C. usinga 0.45 micron PTFE filter and the filtrate collected into clean vials.The clear solution was cooled to −20° C. to induce precipitation. Solidswere isolated for XRPD analysis using filtration or centrifugation.

In some embodiments liquid vapor diffusion was used. In this method aconcentrated stock of compound in solvent is provided. This concentratedstock is transferred to an inner vial that is sealed within a largervial containing anti-solvent. Solids were isolated for XRPD analysisusing filtration or centrifugation.

In some embodiments solid vapor diffusion was used. In this method 5-15mg of sample were weighed into a small (e.g., 3 mL) vial. The vial wasplaced inside a larger vial (e.g., 20 mL) containing 3- to 4 mL of avolatile solvent. The outer vial was then sealed. This assembly was keptat ambient temperature for 7 days, allowing solvent vapor to interactwith the solid, and the resulting product characterized by XRPD.

Unique HM30181 mesylate polymorphisms were characterized by a variety oftechniques, including X-ray powder diffraction (XRPD), NMR, andcalorimetry. These were performed as follows.

XRPD was performed using a Panalytical X'Pert3™ Powder XRPD and on a Sizero-background holder. The 2θ position was calibrated against aPanalytical™ 640 Si powder standard. Details of XRPD used in theexperiments are listed below in Table 20.

TABLE 20 Parameters for Reflection Mode X-Ray wavelength Cu, kα, Kα1(Å): 1.540598, Kα2 (Å): 1.544426 Kα2/Kα1 intensity ratio: 0.50 X-Raytube setting 45 kV, 40 mA Divergence slit Automatic Scan mode ContinuousScan range (°2TH) 3°-40° Step size (°2TH) 0.0131 Scan speed (°/s) 0.16

Differential Scanning calorimetry (DSC) was performed using a TA Q2000™DSC from TA Instruments. Temperature was ramped from ambient temperatureto desired temperature at a heating rate of 10° C./min using N₂ as thepurge gas, with pan crimped (see Table 21).

TABLE 21 Parameters DSC Pan Type Aluminum pan, closed Temperatureambient temperature-300° C. Ramp rate 10° C./min Purge gas N₂In some studies, a cyclic DSC method was used. In such cycling DSCmethods temperature was ramped from ambient to 150° C. at a heating rateof 10° C./min using N₂ as the purge gas, then cooled by 10° C. to 25° C.This temperature cycle repeated twice (see Table 22).

TABLE 22 Parameters DSC Pan Type Aluminum pan, closed Temperature25-150° C. Ramp rate 10° C./min Purge gas N₂ Heat/cool Cycles 2

Thermogravimetric Analysis (TGA) was performed using a TA Q500™ TGA fromTA Instruments. Temperature was ramped from ambient to desiredtemperature at a heating rate of 10° C./min using N₂ as the purge gas,with pan open (see Table 23).

TABLE 23 Parameters TGA Pan Type Platinum plate, open Temperature 300°C. Ramp rate 10° C./min Purge gas N₂ Sample purge flow 15 mL/min Balancepurge flow 25 mL/min

Dynamic Vapor Sorption (DVS) was measured using a SMS (SurfaceMeasurement Systems™) DVS Intrinsic. Parameters for DVS test are listedbelow in Table 24.

TABLE 24 Parameters Values Temperature 25° C. Sample size 10-20 mg Gasand flow rate N₂, 200 mL/min dm/dt 0.002%/min Min. dm/dt stabilityduration 10 min Max. equilibrium time 360 min RH range 40% RH-95% RH-0%RH-95% RH RH step size 10% (0% RH-90% RH) 5% (90% RH-95% RH)

Proton NMR were obtained using a Varian 200M™ NMR in deuterated DMSO(DMSO-d₆).

It should be apparent to those skilled in the art that many moremodifications besides those already described are possible withoutdeparting from the inventive concepts herein. The inventive subjectmatter, therefore, is not to be restricted except in the spirit of theappended claims. Moreover, in interpreting both the specification andthe claims, all terms should be interpreted in the broadest possiblemanner consistent with the context. In particular, the terms “comprises”and “comprising” should be interpreted as referring to elements,components, or steps in a non-exclusive manner, indicating that thereferenced elements, components, or steps may be present, or utilized,or combined with other elements, components, or steps that are notexpressly referenced. Where the specification claims refer to at leastone of something selected from the group consisting of A, B, C . . . andN, the text should be interpreted as requiring only one element from thegroup, not A plus N, or B plus N, etc.

What is claimed is:
 1. A composition comprising a crystalline form ofHM30181 mesylate wherein the crystalline form is polymorph C, andwherein the crystalline form has an X-ray diffraction patterncorresponding to FIG. 42 and has an endotherm at about 159.6° C.
 2. Amethod of inhibiting P-glycoprotein activity, comprising contactingP-glycoprotein with the crystalline form of HM30181 mesylate of claim 1in an amount effective to inhibit an activity of P-glycoprotein.